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J Neurosci. 2019 Mar 29. pii: 0894-18. doi: 10.1523/JNEUROSCI.0894-18.2019. [Epub ahead of print]

Deletion of neuronal GLT-1 in mice reveals its role in synaptic glutamate homeostasis and mitochondrial function.

Author information

1
Department of Drug Design and Pharmacology, University of Copenhagen, 2100 Copenhagen, Denmark.
2
Department of Neurology and the F.M. Kirby Neurobiology Center, Boston Children's Hospital, Boston, Massachusetts, USA 02115.
3
Department of Neuroscience, Norwegian University of Science and Technology, N-7489 Trondheim, Norway.
4
Division of Sleep Medicine, Harvard Medical School, Boston, Massachusetts, USA 02115.
5
Center of Neural Science, New York University, New York, New York, USA 10003.
6
Neuroscience Institute, NYU Langone Medical Center, New York, New York, USA 10016.
7
Department of Neurology and the F.M. Kirby Neurobiology Center, Boston Children's Hospital, Boston, Massachusetts, USA 02115 helle.waagepetersen@sund.ku.dk paul.rosenberg@childrens.harvard.edu.
8
Program in Neuroscience, Harvard Medical School, Boston, Massachusetts, USA 02115.
9
Department of Drug Design and Pharmacology, University of Copenhagen, 2100 Copenhagen, Denmark helle.waagepetersen@sund.ku.dk paul.rosenberg@childrens.harvard.edu.

Abstract

The glutamate transporter GLT-1 is highly expressed in astrocytes but also in neurons, primarily in axon terminals. We generated a conditional neuronal GLT-1 knockout using synapsin 1-Cre (synGLT-1 KO) to elucidate the metabolic functions of GLT-1 expressed in neurons, here focusing on the cerebral cortex. Both synaptosomal uptake studies and electron microscopic immunocytochemistry demonstrated knockdown of GLT-1 in the cerebral cortex in the synGLT-1 KO mice. Aspartate content was significantly reduced in cerebral cortical extracts as well as synaptosomes from cerebral cortex of synGLT-1 KO compared to control littermates. 13C-Labelling of tricarboxylic acid cycle intermediates originating from metabolism of [U-13C]-glutamate was significantly reduced in synGLT-1 KO synaptosomes. The decreased aspartate content was due to diminished entry of glutamate into the TCA cycle. Pyruvate recycling, a pathway necessary for full glutamate oxidation, was also decreased. ATP production was significantly increased, despite unaltered oxygen consumption, in isolated mitochondria from the synGLT-1 KO. Density of mitochondria in axon terminals and peri-synaptic astrocytes were increased in the synGLT-1 KO. Intramitochondrial cristae density of synGLT-1 KO mice was increased suggesting increased mitochondrial efficiency, perhaps in compensation for reduced access to glutamate. SynGLT-1 KO synaptosomes exhibited an elevated oxygen consumption rate when stimulated with veratridine, despite a lower baseline oxygen consumption rate in the presence of glucose. GLT-1 expressed in neurons appears to be required to provide glutamate to synaptic mitochondria and is linked to neuronal energy metabolism and mitochondrial function.SIGNIFICANCE STATEMENTAll synaptic transmitters need to be cleared from the extracellular space after release, and transporters are used to clear glutamate released from excitatory synapses. GLT-1 is the major glutamate transporter, and most GLT-1 is expressed in astrocytes. Only 5-10% is expressed in neurons, primarily in axon terminals. The function of GLT-1 in axon terminals remains unknown. Here, we used a conditional knockout approach to investigate the significance of the expression of GLT-1 in neurons. We found multiple abnormalities of mitochondrial function suggesting impairment of glutamate utilization by synaptic mitochondria in the neuronal GLT-1 knockout. These data suggest that GLT-1 expressed in axon terminals may be important in maintaining energy metabolism and biosynthetic activities mediated by presynaptic mitochondria.

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